Study of surface treated historical materials

Import 05/08/2014Diplomová práce se zabývá obecným studiem povrchových úprav technikou plátkového zlacení na historických předmětech, zejména obrazových rámech. V teoretické části práce pojednává o historii a principech zhotovování technologií zlacení povrchu různých materiálů od minulosti k současnosti. Zřetel je kladen na dřevěné obrazové rámy a jejich části. Cílem práce je průzkum složení povrchových úprav a to jak složení slitiny použitého plátkového kovu, tak složení pojiv a podkladové vrstvy. Experimentální část je zaměřena na průzkum povrchu historických předmětů pomocí skenovací elektronové mikroskopie a analýzu typu syntetických pryskyřic rámů pomocí spektroskopických a chromatografických metod.This thesis deals with the general study of finishes leaf gilding technique on historical subjects, especially picture frames. The theoretical part deals with the history and principles of making technology gilding the surface of various materials from past to present. Consideration shall be given to the wooden picture frames and parts. The aim of the work is investigation of the finishes in both the composition of the alloy used leaf metal and binder composition and the underlying layer. The experimental part will focus on exploration of historical objects surface using scanning electron microscopy and analysis of the type of synthetic resin frames using chromatographic methods.636 - Katedra materiálového inženýrstvívýborn

AKT activation by N-cadherin regulates beta-catenin signaling and neuronal differentiation during cortical development

Background: During cerebral cortical development, neural precursor-precursor interactions in the ventricular zone neurogenic niche coordinate signaling pathways that regulate proliferation and differentiation. Previous studies with shRNA knockdown approaches indicated that N-cadherin adhesion between cortical precursors regulates β-catenin signaling, but the underlying mechanisms remained poorly understood. Results: Here, with conditional knockout approaches, we find further supporting evidence that N-cadherin maintains β-catenin signaling during cortical development. Using shRNA to N-cadherin and dominant negative N-cadherin overexpression in cell culture, we find that N-cadherin regulates Wnt-stimulated β-catenin signaling in a cell-autonomous fashion. Knockdown or inhibition of N-cadherin with function-blocking antibodies leads to reduced activation of the Wnt co-receptor LRP6. We also find that N-cadherin regulates β-catenin via AKT, as reduction of N-cadherin causes decreased AKT activation and reduced phosphorylation of AKT targets GSK3β and β-catenin. Inhibition of AKT signaling in neural precursors in vivo leads to reduced β-catenin-dependent transcriptional activation, increased migration from the ventricular zone, premature neuronal differentiation, and increased apoptotic cell death. Conclusions: These results show that N-cadherin regulates β-catenin signaling through both Wnt and AKT, and suggest a previously unrecognized role for AKT in neuronal differentiation and cell survival during cortical development

Additional file 2: Figure S2. of Afadin controls cell polarization and mitotic spindle orientation in developing cortical radial glia

Expression of adherens junction-associated proteins in control and Afadin-deleted dorsal forebrains at E13.5. (A, B) Western blot analysis of E13.5 dorsal forebrain extracts show a strong reduction of Afadin expression in mutant, but not of intercellular junctions-associated proteins α-catenin, N-cadherin, β-catenin, p120catenin, Z0-1, or any cadherin when compared to control (mean ± s.d. Unpaired t-test.; n = 3 to 8 animals). (TIF 2373 kb

The Lis1–Nde1 complex is essential for determining cerebral cortical size, shape and lamina structures

() Dying at birth, the brains of Nde1Lis1 mutant mice were dramatically reduced (arrows). A more pronounced size reduction of the cerebral hemispheres was observed. () Histological analysis showed that the neocortex of the Nde1Lis1 mutant was severely disorganized, lacked the normal MZ and any other cortical layers. () Immunohistological analyses with layer-specific markers showed that in the Nde1Lis1 cortex, cells belonging to superficial and middle cortical layers (marked by Cux1 and Foxp1 immunoreactivity, respectively) were greatly reduced, positioned randomly and often formed heterotopia in deeper cortical regions. In contrast, deep layer maker Tbr1 highlighted cells in the superficial cortex, suggesting grossly inverted cortical layers in the mutant. E, Nde1; L, Lis1. Bar: 200 µm () Rostral to caudal length (L1), medial to lateral length (L2) and cortical thickness (V) of Nde1Lis1 cortex were measure and compared with those of the Nde1 control at P0. The mutant cortex was less than 40% of the controls in L1 and L2, but only reduced by 20% in thickness (V).<p><b>Copyright information:</b></p><p>Taken from "Lis1–Nde1-dependent neuronal fate control determines cerebral cortical size and lamination"</p><p></p><p>Human Molecular Genetics 2008;17(16):2441-2455.</p><p>Published online 10 May 2008</p><p>PMCID:PMC2486443.</p><p>© 2008 The Author(s).</p

Morphology and cytoarchitectural defects of Nde1Lis1 progenitors underlie the abnormal fate control

() Immunostaining the cell body of metaphase progenitors with a phosphorylated vimentin antibody 4A4 showed moderately reduced apical staining of VZ progenitors in the Nde1, Nde1Lis1 mutants and a severe decrease in 4A4 immunoreactivity along the ventricular surface in the Nde1Lis1 mutant (<p><b>Copyright information:</b></p><p>Taken from "Lis1–Nde1-dependent neuronal fate control determines cerebral cortical size and lamination"</p><p></p><p>Human Molecular Genetics 2008;17(16):2441-2455.</p><p>Published online 10 May 2008</p><p>PMCID:PMC2486443.</p><p>© 2008 The Author(s).</p

Overproduction of PP neurons and lack of preplate splitting in the Nde1 Lis1 cortex

() Preplate neurons were labeled by CSPG antibody in red and cortical plate neurons were labeled with DCX antibody in green. A significant increase in CSPG was detected throughout the entire Nde1Lis1 cortex before its size was reduced at E11.5. () The cortex of Nde1Lis1 mutant was thinner at E12.5, but their CSPG positive zone was broadened (arrows). () At E13.5, while Nde1, Nde1 and Nde1Lis1 embryos all showed well separated preplate (indicated by yellow arrows), no splitting of the preplate could be detected in the Nde1Lis1 cortex. In addition to greatly increased CSPG positive preplate neurons (in red), increases of DCX positive young cortical plate neurons (in green) was also detectable. E, Nde1; L, Lis1. Bar: 100 µm.<p><b>Copyright information:</b></p><p>Taken from "Lis1–Nde1-dependent neuronal fate control determines cerebral cortical size and lamination"</p><p></p><p>Human Molecular Genetics 2008;17(16):2441-2455.</p><p>Published online 10 May 2008</p><p>PMCID:PMC2486443.</p><p>© 2008 The Author(s).</p

Reduction of VZ progenitors at the onset of corticogenesis by Lis1–Nde1 double deficiency

() The size of telencephalic vesicles (indicated by dashed blue circles) of the Nde1Lis1 mutant appeared close to normal before E11, but was significantly smaller by E13 compared to that of their littermates. () Correlated with the thinning of cerebral cortex at E12.5, the thickness of cortical VZ and the number of S-phase neural progenitors examined by BrdU transient labeling (in red), was decreased significantly in the Nde1Lis1 mutant. < 0.001. E, Nde1; L, Lis1. Bar: 100 µm. () Reduced neural progenitors in the Nde1Lis1 mutant cortex was indicated by reduced immunostaining of the glutamate transporter GLAST (in red), a marker of radial glial progenitors. Bar: 100 µm. () Substantial cell death was detected in the cerebral cortex of the Nde1Lis1 mutant by TUNEL staining (in green). In contrast, very few TUNEL positive cells were detected in the Nde1 and Nde1Lis1 controls. Bar: 100 µm. () Majority (over 80%) of TUNEL positive cells (in green) in the Nde1Lis1 mutant were newborn post mitotic neurons in the intermediate zone (IZ) and the cortical plate (CP) and expressed DCX (in red). Bar: 100 µm. () Cell-cycle exit profiles of Nde1Lis1progenitors. Pregnant females were given single does of BrdU at E12 and were analyzed 18 h later. Brain sections were stained with antibody to BrdU (in green) and to Ki67 (in red). Cells that exit the cell cycle are counted as those that are positive for BrdU but negative for Ki67, and presented as the percentage of total BrdU positive cells. Approximately 67% of cells labeled by BrdU at E12 became non-progenitor cells in the Nde1Lis1 cortex by E13, while only 25–28% of the Nde1Lis1 or Lis1 progenitors left the cell cycle (<p><b>Copyright information:</b></p><p>Taken from "Lis1–Nde1-dependent neuronal fate control determines cerebral cortical size and lamination"</p><p></p><p>Human Molecular Genetics 2008;17(16):2441-2455.</p><p>Published online 10 May 2008</p><p>PMCID:PMC2486443.</p><p>© 2008 The Author(s).</p

Overproduction of C–R cells and enhanced Reelin signaling in the Nde1Lis1 cortex

() Over production of Calretinin positive C–R cells (in red) was seen in the Nde1Lis1 mutant. Significantly increased Calretinin positive cells were observed in the Nde1Lis1 cortex at E12.5. E, Nde1; L, Lis1. Bar: 200 µm. ( and ) Over-production and abnormal positioning of Reelin secreting C–R cells in the Nde1Lis1 mutant. Cortical sections were immunostained with two monoclonal antibodies to Reelin in red (CR50 and G10), and co-stained with an antibody to DCX in green and Hoechst in blue. The Nde1Lis1 mutant showed dramatic increase and mis-localization of Reelin positive C–R cells throughout the entire course of corticogenesis starting from E11.5. Bar: 200 µm. () Immunoblotting analysis of Reelin protein levels in developing cerebral cortex. The cerebral cortices of E13.5 and E15.5 embryos were dissected and their total proteins extracts were analyzed on a 7.5% SDS–PAGE, followed by immunoblotting with an antibody to Reelin (G10). Loading is normalized by total protein amount and by immunoblotting with an antibody to tubulin. Bands on immunoblots were analyzed using Quantify One. Over 5-fold increase in Reelin protein (both 400 and 180 kDa bands) was detected in the Nde1Lis1 mutant cortex over wild-type controls. () Down-regulation of Dab1 by increased Reelin signaling was observed in the Nde1Lis1 mutant. Immunoblotting analysis was performed with protein extracts from the cortex of E15.5 embryos with antibodies to Dab1 and DCX. Compared to wild-type and Nde1Lis1 mutant, a significant decrease in the Dab1 protein level was detected in the Nde1Lis1cortex, suggesting that the overproduced Reelin in the mutant were active and could elicited Dab1 degradation.<p><b>Copyright information:</b></p><p>Taken from "Lis1–Nde1-dependent neuronal fate control determines cerebral cortical size and lamination"</p><p></p><p>Human Molecular Genetics 2008;17(16):2441-2455.</p><p>Published online 10 May 2008</p><p>PMCID:PMC2486443.</p><p>© 2008 The Author(s).</p

CDH11 is differentially expressed in human glioma vs. normal brain, and overexpression confers worsened prognosis.

<p>(<b>A</b>) Kaplan-Meier Survival Plot for samples with differential <i>CDH11</i> gene expression reveal that glioma patients with overexpression (red curve) have a worse prognosis than patients with intermediate expression. Log-rank P-value for Up-Regulated vs. Intermediate = 3.8535E-6. Data from NCI 2005<<a href="http://rembrandt.nci.nih.gov" target="_blank">http://rembrandt.nci.nih.gov</a>>. (<b>B</b>) <i>CDH11</i> expression is predictive for survival in GBM; fold-change 3.6, P = 0.005 by T-test (Data from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Nutt1" target="_blank">[38]</a>). (<b>C</b>) <i>CDH11</i> expression is higher in GBM vs. mixed glioma or oligodendroglial tumors; fold-change = 1.8, P = 5.83×10⁻⁴ by T-Test (Data from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Liang1" target="_blank">[39]</a>). (<b>D</b>) <i>CDH11</i> expression is higher in GBM vs. neural stem cells. Fold change = 7.8, P = 8.92×10⁻⁴ (Data from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Lee1" target="_blank">[40]</a>). (<b>E</b>) <i>CDH11</i> is relatively overexpressed in GBM vs. normal brain tissue. TCGA glioblastoma dataset analysis shows 2.9 fold overexpression of CDH11 (P = 1.14×10^−10, T-Test). GBM vs. Normal in (Bredel <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Bredel1" target="_blank">[41]</a>): 2.2 fold increase; P = 0.018 (T-test). GBM vs. Normal (Sun <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Sun1" target="_blank">[42]</a>): 2.1 fold increase (P = 1.14×10⁻¹⁶, T-test). GBM vs. normal (Liang <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Liang1" target="_blank">[39]</a>): 3.6 fold increase (P = 0.005, T-test). GBM vs. WM (Shai <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Shai1" target="_blank">[43]</a>): 2.5 fold overexpression compared to white matter (P = 9.75×10⁻⁸; T-test). (<b>F–I</b>) Relative expression of cadherin genes in GBM vs. normal from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Sun1" target="_blank">[42]</a> (<b>F</b>), <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Liang1" target="_blank">[39]</a> (<b>G</b>), <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Bredel1" target="_blank">[41]</a> (<b>H</b>), and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070962#pone.0070962-Lee1" target="_blank">[40]</a> (<b>I</b>) reveals that CDH11 (boxed in red) is differentially regulated in GBM vs. normal across multiple datasets. (<b>J–M</b>) Immunoperoxidase staining for CDH11 expression in human GBM tissue show heterogenous expression patterns, with enrichment of staining in tumor cells adjacent to tumor vessels (arrows). Cells surrounding vessels of varying sizes from small (<b>J</b>) to large vessels (<b>K</b>) express Cdh11. As typical for GBM, the tumor histoarchitecture is highly varied, with tissue showing variable necrosis and marked heterogeneity of cellularity (notably (<b>J</b>) and (<b>K</b>)). Bar = 100 µm.</p

Overexpression of CDH11 in cortical VZ precursors causes premature exit from the VZ and neuronal differentiation.

<p>(<b>A</b>) E13.5 cortical precursors were electroporated in utero using either pCDNA control (N = 4) (top) or with pCAG-Cdh11 expression plasmid along with pCAG-EGFP plasmid (N = 4) and analyzed at E14.5. Electroporated cells were identified with antibody staining against GFP and sections counterstained with the DNA dye DAPI (pseudocolored blue). To quantify changes in cortical positioning of electroporated cells, ten equal sized bins were drawn over each image. Each white dot corresponds with the soma of an electroporated cell. Bar = 100 µm. (<b>B</b>) The fraction of the total GFP+ cells in each of the ten bins was then graphed for the two experimental conditions. Brackets indicate 1 SEM. N = 4 brains (PCDNA), 3 brains (Cdh11). (<b>C</b>) Sections were stained for radial glial marker Pax6, intermediate progenitor marker Tbr2, and neuronal marker Tbr1. Electroporated cells are pseudocolored green, and the respective antigens, red. Bar = 50 µm. The dot plots highlight the cell bodies of electroporated cells, with red representing electroporated cells that express the marker of interest and green indicating electroporated cells that do not express the marker. (<b>D</b>) Histograms represent fraction of total electroporated cells found in each brain region, showing the fraction of cells that express each marker after electroporation (red/(red+green)), showing premature neuronal differentiation. For Pax 6, Cdh11 vs. control (N = 4 brains for each, **P = 0.0072), Tbr2 (N = 3 for Cdh11, N = 4 for pcDNA control, *P = 0.077), and Tbr1 (N = 4 for each, *P = 0.0071). Unpaired T-test.</p